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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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人血浆中mbl-masp复合物的纯化与分离
, PP. 1373-1377
Keywords: 甘露聚糖结合凝集素,甘露聚糖结合凝集素相关丝氨酸蛋白酶,亲和层析,凝胶过滤
Abstract:
目的从人血浆中分离纯化甘露聚糖结合凝集素(mbl)和mbl相关丝氨酸蛋白酶(masp)。方法用非活化sepharose4b两步亲和层析纯化mbl-masp复合物,再以sephacryls-300柱凝胶过滤将mbl和masp分离。在纯化mbl-masp复合物的整个过程中,除凝胶过滤缓冲液外,均加入丝氨酸蛋白酶抑制剂苯甲磺酰氟(pmsf)和金属蛋白酶抑制剂1,10-邻二氮杂菲(1,10-phenanthroline),并控制温度在4℃。结果获得高度纯化的mbl和酶原形式的masp。sds-page和westernblotting表明所纯化的mbl相对分子质量为28000和32000肽链构成的功能性多聚体;配体结合测定和酵母菌凝集试验证实所纯化的mbl具有生物学活性。结论建立了一种比较简便的同时分离纯化mbl和masp酶原的方法。
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